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泰山医学院毕业设计（论文）

题目：蜂胶黄酮抗H2O2诱导PC12细胞

凋亡机制的研究

院（部）系

药学院

所学专业药学

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2013年 6 月 18 日

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20 年月日

摘 要

目的 观察蜂胶黄酮对过氧化氢（H 2O 2）诱导大鼠肾上腺嗜铬细胞瘤细胞（PC 12）凋亡的影响及机制。

方法 培养PC 12细胞，取对数生长期细胞分为五组，空白对照组、模型组、蜂胶黄酮高、中、低剂量组，剂量分别为200mg/L 、100 mg/L 、50 mg/L.药物预处理2h 后，孵育H 2O 2（140μmol/L）24h 诱导过氧化损伤。TUNEL 试剂盒检测原位细胞凋亡,流式细胞仪检测细胞内活性氧水平以及细胞周期，ELISA 检测细胞内Caspase-3蛋白含量。

结果 TUNEL 细胞凋亡染色显示, H 2O 2组与空白对照组比较染色明显加深,而蜂胶黄酮组染色明显变浅，说明蜂胶黄酮能对抗H 2O 2诱导细胞凋亡；周期结果显示H 2O 2组处于G0/G1细胞明显增多，而处于G2/M 、S 期细胞明显减少，细胞增殖降低，而蜂胶黄酮增加S 期细胞促进细胞增殖；与空白对照组相比H 2O 2组活性氧水平、细胞内Caspase-3含量明显增高，而蜂胶黄酮各剂量组活性氧水平降低，Caspase-3含量减少。

结论 蜂胶黄酮对H 2O 2诱发PC 12神经细胞凋亡有显着的抑制作用，其机制可能其影响细胞周期、清除氧自由基、降低凋亡因子Caspase-3有关。

关键词 蜂胶黄酮；PC 12细胞；H 2O 2;Caspase-3；细胞凋亡

Abstract

Objective :

To observe the protection ofpropolis flavonoidson rat with injuried pheochromocytomacells(PC 12) induced by?hydrogen peroxide(H 2O 2) and to explore its possible mechanism.

Methods:Culture?PC 12 cells,the cells inthe logarithmic growth phase weredivided into five groups,blank control group,model group,propolis flavoneof high,medium and low dose group,the dose of 200mg/L,100 mg/L,50 mg/L.After 2h 0fPharmacological preconditioning,the cellswere incubated with H 2O 2(140 μ mol/L)for24hto induceoxidative damage.TUNEL Kitis used fordetermining Cell apoptosis,Flow cytometry was used to detect the intracellular ROS level and cell cycle,ELISA is used for determining the concentration of protein Caspase-3 in cells.

Results:Apoptosis of TUNEL cells staining, H 2O 2 group compared with the control group, staining was deepened, and the propolis flavone group was significantly lighter, that propolis flavonoids can antagonize the apoptosis induced by H 2O 2 Cycle showed that H 2O 2 group in G0/G1 cells were increased, and in the G2/M,S phase cells decreased significantly, reduced cell proliferation, and propolis flavonoids increased S phase cells promoting cell proliferation; compared with the blank control group, caspase-3 in H 2O 2group, the levels of reactive oxygen species in cells increased significantly, while the propolis flavone in all dose groups decrease the levels of reactive oxygen species, reduced caspase-3 content.

Conclusion : Propolis flavonoids on H 2O 2-induced PC 12 cells damage a significant protective effect,The mechanism may be its effect on cell cycle, scavenging oxygen free radicals, decrease the apoptosis related factor caspase-3.

Key words : Propolis; flavonoids; PC 12 cells; H 2O 2; apoptosis